菊花CmTPS1like基因的克隆及表达特性

Cloning and expression characteristics of CmTPS1like gene in Chrysanthemum morifolium

[目的]本文旨在克隆菊花β-石竹烯合成酶同源基因(CmTPS 1like)并探讨其参与蚜虫取食和茉莉酸甲酯(MeJA)处理应答表达的机制。[方法]以菊花‘神马’为试验材料,采用高保真PCR及染色体位移法分别克隆CmTPS 1like基因全长及上游启动子序列,并采用RT-qPCR分析CmTPS 1like基因的组织特异性表达以及响应蚜虫取食和MeJA处理的表达模式。[结果]CmTPS 1like基因开放阅读框(ORF)共有1644个碱基,编码548个氨基酸,且与菊科植物野菊同源性最高(95.26%)。CmTPS 1like基因上游启动子区包括2个MeJA响应元件、5个MYC结合元件、2个MYB结合元件、5个光响应元件等。CmTPS 1like基因在茎中表达量最高,叶和根中表达量次之,花中表达量最低。蚜虫取食12和24 h后,菊花叶片中CmTPS 1like基因的表达水平上调。随着MeJA处理时间的增加(3~24 h),CmTPS 1like基因被持续诱导表达。[结论]CmTPS 1like基因在菊花‘神马’茎中表达量最高,其表达受蚜虫取食和MeJA处理的影响。

[Objectives]The purpose of this research was to clone Chrysanthemum morifoliumβ-caryophyllene synthesis enzyme homologous gene CmTPS 1 like and to explore the response mechanism of aphid feeding and methyl jasmonate(MeJA)treatment.[Methods]The full-length and the partial sequence of promoter of CmTPS 1 like was obtained by high fidelity PCR and chromosome walking using chrysanthemum‘Jinba’as material.The relative expression levels of CmTPS 1 like in different tissues of chrysanthemum‘Jinba’were detected by RT-qPCR.The expression profiles of CmTPS 1 like under aphid infestation and MeJA treatment were analyzed.[Results]The open reading frame(ORF)of CmTPS 1 like had 1644 bp encoding 548 amino acids.Phylogenetic analysis showed that it had the closest relationship with TPS1 from Chrysanthemum indicum with an identity of 95.26%.The promoter sequence of CmTPS 1 like contained two MeJA responsive elements,five MYC binding elements,two MYB binding elements,five light responsive elements,etc..CmTPS 1 like had the highest expression level in stem,followed by leaves and roots,the lowest in flowers.After aphid feeding,the expression level of CmTPS 1 like at 12 and 24 h increased.Moreover,the expression of CmTPS 1 like was induced by MeJA treatment(3-24 h).[Conclusions]The expression level of CmTPS 1 like gene was the highest in the stem.The expression of CmTPS 1 like was regulated by aphid feeding and MeJA treatment.